脾虚痰湿型肥胖糖尿病胰岛素抵抗大鼠病证结合模型的建立

Establishment of insulin resistance diabetes and obesity rat models with syndrome of phlegm-dampness due to spleen deficiency

目的:建立一种病证结合的脾虚痰湿型肥胖糖尿病胰岛素抵抗大鼠模型。方法:60只SD大鼠按体质量随机分为正常对照(A)组和模型组,模型组又分为肥胖糖尿病(B)组、脾虚痰湿型肥胖糖尿病(C)组和脾虚痰湿肥胖糖尿病加中药健脾化湿方治疗(D)组。模型组复制肥胖糖尿病胰岛素抵抗大鼠疾病模型,C组和D组复制脾虚痰湿型肥胖糖尿病胰岛素抵抗大鼠模型,D组给予健脾化湿方干预4周。观察各组大鼠的一般行为学变化、毛色,大便、体质量、摄食量、饮水量、肛温、游泳耐力、计算脾虚积分;测定大鼠的空腹血糖(FPG)、空腹胰岛素(Fins),计算胰岛素抵抗指数(HOMA-IR)、胰岛素敏感指数(ISI);口服葡萄糖耐量试验(OGTT)、总胆固醇(TC)、甘油三脂(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、游离脂肪酸(FFA);观察肝脏病理变化。并采用健脾化湿方进行反证。结果:高脂饲料喂养8周后,模型组平均体质量超过A组(P<0.05);STZ注射72h后,模型组大鼠FPG、Fins、HOMA-IR明显高于A组(P<0.05),ISI明显低于A组(P<0.01);模型组大鼠OGTT各点血糖值均高于A组(P<0.05)。经过4周脾虚痰湿阶段造模后,大鼠表现为体质量下降、饮食量、饮水量明显减少,游泳耐力显著下降,被毛油腻,倦怠,懒动,蜷缩扎堆,大便溏,脾虚积分显著增加(P<0.05,P<0.01),肛温无明显变化;TC、LDL-C、FFA升高(P<0.05,P<0.01)。经过中药健脾化湿方干预后,大鼠出现体质量下降,饮食量、饮水量下降,游泳耐力增加,倦怠、乏力,大便溏症状缓解,脾虚积分下降(P<0.05,P<0.01);大鼠FPG、Fins、HOMA-IR、ISI明显改善(P<0.05,P<0.01);血脂中TC、TG、LDL-C、FFA降低(P<0.05,P<0.01);肝脏病理结果显示,B、C组伴有不同程度的脂肪肝,D组脂肪肝程度减轻。结论:采用高脂饮食联合小剂量STZ可复制出肥胖糖尿病胰岛素抵抗大鼠疾病模型,采用饮食不节+劳倦过度+苦寒攻下复合法可成功建立脾虚痰湿型大鼠证候模型。

Objective： To establish the rat models of insulin resistance diabetes and obesity with syndrome of phlegmdampness due to spleen deficiency. Methods： Sixty SD rats were randomly divided into control group（A） and model group. Model group was divided into model control group（B）, phlegm-dampness due to spleen deficiency group（C） and TCM group（D）. Model group was replicated the model of obese diabetic rats with insulin resistance. Phlegm-dampness due to spleen deficiency group and TCM group were replicated phlegm-dampness due to spleen deficiency type of obese diabetic model with insulin resistance disease. The TCM groupwas given intervention of invigorating spleen for eliminating dampness decoction for 4 weeks. The general behavior changes of the rats including hair color, stool, body weight, food consumption, water intake, rectal temperature, swimming endurance and spleen deficiency integral calculation were observed. Fasting plasma glucose（FPG）, fasting insulin（Fins）, oral glucose tolerance test（OGTT）, total cholesterol（TC）, triglyceride（TG）, high density lipoprotein cholesterol（HDL-C）, low density lipoprotein cholesterol（LDL-C） and free fatty acid（FFA） were observed in rats. Insulin resistance index（HOMA-IR） and insulin sensitivity index（ISI） were calculated. Pathological changes of the liver were also observed. Invigorating spleen for eliminating dampness decoction was used to provide counterevideace. Results： After given high-fat diet for 8 weeks, average body mass in high-fat diet group was higher than normal food group（P0.05）. After given STZ injection 72 h, the FPG, Fins, and HOMA-IR in rats of model group were significantly higher than the control group（P0.05）. ISI was significantly lower than the control group（P0.01）. OGTT glucose values at each time in model group were higher than the control group（P0.05）. After the stage of producing phlegm-dampness due to spleen deficiency model, model rats showed that body weight was decreased. Food consumption and water intake were significantly decreased. Swimming endurance was significantly reduced. Furthermore, rats showed greasy hair, fatigue, unwilling to exercise, curling get together, loose stools and increasing in spleen deficiency integral（P0.05, P0.01）. Rectal temperature did not change significantly. TC, LDL-C and FFA increased（P0.05, P0.01）. After given intervention of invigorating spleen for eliminating dampness decoction, rats showed that body weight, food consumption and water intake were decreased. Swimming endurance was increased. Symptoms of fatigue, weakness and loose stools were alleviated. Spleen deficiency integral was decreased（P0.05, P0.01）. Rats of FPG, Fins, HOMA- IR and ISI were significantly ameliorated（P0.05, P0.01）. Lipid（TC, TG, LDL-C） and FFA were declined（P0.05, P0.01）. Liver pathology results showed different degrees of fatty liver in B and C group, and phlegm-dampness due to spleen deficiency group. The degree of fatty liver was reduced in D group. Conclusion： The method of using high-fat diet combined with low-doses streptozotocin can replicate a model of obese diabetic insulin resistance rats. Using complex methods of improper diet, over exertion, bitter cold and purgation can be successfully established a model of Spleen deficiency and dampness accumulation.